1. Field of the Invention
The present invention generally concerns optical tweezers, microfluidics, flow cytometry, biological Micro Optical Electro Mechanical Systems (Bio-MOEMS), Laguerre-Gaussian mode emissions from Vertical Cavity Surface Emitting Lasers (VCSELs), cell cytometry and microfluidic switches and switching.
The present invention particularly concerns the sorting of microparticles in fluid, thus a “microfluidic sorting device”; and also the directed movement, particularly for purposes of switching, of microparticles based on the transference of momentum from photons impinging on the microparticles, ergo “photonic momentum transfer”.
2. Description of the Prior Art
2.1 Background to the Functionality of the Present Invention
In the last several years much attention has been paid to the potential for lab-on-a-chip devices to significantly enhance the speed of biological and medical research and discovery. See P. Swanson, R. Gelbart, E. Atlas. L. Yang, T. Grogan, W. F. Butler, D. E. Ackley, and C. Sheldon. “A fully multiplexed CMOS biochip for DNA analysis,” Sensors and Actuators B 64, 22–30 (2000). See also M. Ozkan, C. S. Ozkan, M. M. Wang, O. Kibar, S. Bhatia, and S. C. Esener, “Heterogeneous Integration of Biological Species and Inorganic Objects by Electrokinetic Movement,” IEEE Engineering in Medicine and Biology, in press.
The advantages of such bio-chips that have been demonstrated so far include the abilities to operate with extremely small sample volumes (on the order of nanoliters) and to perform analyses at much higher rates than can be achieved by traditional methods. Devices for study of objects as small as DNA molecules to as large as living cells have been demonstrated. See P. C. H. Li and D J, Harrison, “Transport, Manipulation, and Reaction of Biological Cells On-Chip Using Electrokinetic Effects,” Anal. Chem. 69, 1564–1569 (1997).
One important capability for cell research is the ability to perform cell sorting, or cytometry, based on the type, size, or function of a cell. Recent approaches to micro-cytometry have been based on electrophoretic or electro-osmotic separation of different cell types. See A. Y. Fu, C. Spence, A. Scherer, F. H. Arnold, and S. R Quake, “A microfabricated fluorescence-activated cell sorter,” Nature 17.1109–1111 (1999).
2.2 Scientific Background to the Structure of the Device of the Present Invention
The present invention will be seen to employ optical tweezers. See A. Ashkin, J. M. Dziedzic, J. E. Bjorkholm, and S. Chu, “Observation of a single-beam gradient force optical trap for dielectric particles;” Opt. Lett. 11, 288–291) (1986).
The present invention will also be seen to employ micro-fabricated fluidic channels. See H.-P. Chou, C. Spence. A. Scherer. and S. Quake, “A microfabricated device for sizing and sorting DNA molecules,” Proc. Natl. Acad. Sci. USA 96 11–13 (1999).
In previous demonstrations of the optical manipulation of objects through defined fluidic channels, photonic pressure was used to transport cells over the length of the channels. See T. N. Buican M. J. Smyth, H. A. Crissman, G. C. Salzman, C. C. Stewart, and J. C. Martin, “Automated single-cell manipulation and sorting by light trapping.” Appl. Opt, 26, 3311–5316 (1987). The device of the present invention will be seen to function oppositely.
2.3 Engineering, and Patent, Background to the Structure of the Device of the Present Invention
There are many existing (i) bio-chip (lab-on-a-chip) technologies, and (ii) microfluidic technologies. Most of these technologies use electrical or mechanical force to perform switching within the channels. The present invention is unique in that optics (as generate photonic pressure, or radiation pressure) is used to perform switching—particularly of small particles flowing in microfluidic channels.
2.3.1 Background Patents Generally Concerning Optical Tweezing and Optical Particle Manipulation
The concept of using photonic pressure to move small particles is known. The following patents, all to Ashkin, generally deal with Optical Tweezers. They all describe the use of optical “pushing” and optical “trapping” forces, both of which are used in the present invention. These patents do not, however, teach or suggest such use of optical forces in combination with microfluidics as will be seen to be the essence of the present invention.
U.S. Pat. No. 3,710,279 to Askin, assigned to Bell Telephone Laboratories, Inc. (Murray Hill, N.J.), for APPARATUSES FOR TRAPPING AND ACCELERATING NEUTRAL PARTICLES concerns a variety apparatus for controlling by radiation pressure the motion of particle, such as a neutral biological particle, free to move with respect to its environment. A subsequent Askin patent resulting from a continuation-in-part application is U.S. Pat. No. 3,808,550.
Finally, U.S. Pat. No. 4,893,886 again to Ashkin, et al., assigned to American Telephone and Telegraph Company (New York, N.Y.) and AT&T Bell Laboratories (Murray Hill, N.J.), for a NON-DESTRUCTIVE OPTICAL TRAP FOR BIOLOGICAL PARTICLES AND METHOD OF DOING SAME, concerns biological particles successfully trapped in a single-beam gradient force trap by use of an infrared laser. The high numerical aperture lens objective in the trap is also used for simultaneous viewing. Several modes of trapping operation are presented.
2.3.2 Patents Showing Various Conjunctions of Optical Tweezing/Optical Manipulation and Microfluidics/Microchannels
U.S. Pat. No. 4,887,721 to Martin, et al., assigned to Bell Telephone Laboratories, Inc. (Murray Hill, N.J.), for a LASER PARTICLE SORTER, concerns a method and apparatus for sorting particles, such as biological particles. A first laser defines an optical path having an intensity gradient which is effective to propel the particles along the path but which is sufficiently weak that the particles are not trapped in an axial direction. A probe laser beam interrogates the particles to identify predetermined phenotypical characteristics of the particles. A second laser beam intersects the driving first laser beam, wherein the second laser beam is activated by an output signal indicative of a predetermined characteristic. The second laser beam is switchable between a first intensity and a second intensity, where the first intensity is effective to displace selected particles from the driving laser beam and the second intensity is effective to propel selected particles along the deflection laser beam. The selected particles may then be propelled by the deflection beam to a location effective for further analysis.
The described particle propulsion means of Martin, et al. concerns (i) the suspension of particles by fluidics and (ii) the use of an optical pushing beam to move particles around in a cavity. The application of sorting—as is performed by certain apparatus of the present invention—is also described. However, the present invention is distinguished over U.S. Pat. No. 4,887,721 for SORTING IN MICROFLUIDICS to Martin, et al. because this patent teaches the use of optical beams to do all particle transport, while the present invention uses optical beams only for switching, with transport accomplished by microfluidic flow. In the apparatus of U.S. Pat. No. 4,887,721 a single beam pushes a particle along from one chamber to the next. It will soon be seen that in the various apparatus of the present invention continuous water flow serves to move the particles around, and optics is only used as the switch. This is a much more efficient use of photons and makes for a faster throughput device.
The Martin, et al. patent also describes (i) sensing particles by optical means, and (ii) act on the results of the sensing so as to (iii) manipulate the particles with laser light. Such optical sensing is fully compatible with the present invention.
Also involving both (i) fluidics and, separately, (ii) optical manipulation is U.S. Pat. No. 5,674,743 to Ulmer, assigned to SEQ, Ltd. (Princeton, N.J.), for METHODS AND APPARATUS FOR DNA SEQUENCING. The Ulmer patent concerns a method and apparatus for automated DNA sequencing. The method of the invention includes the steps of: a) using a processive exonuclease to cleave from a single DNA strand the next available single nucleotide on the strand; b) transporting the single nucleotide away from the DNA strand; c) incorporating the single nucleotide in a fluorescence-enhancing matrix; d) irradiating the single nucleotide to cause it to fluoresce; e) detecting the fluorescence; f) identifying the single nucleotide by its fluorescence; and g) repeating steps a) to f) indefinitely (e.g., until the DNA strand is fully cleaved or until a desired length of the DNA is sequenced). The apparatus of the invention includes a cleaving station for the extraction of DNA from cells and the separation of single nucleotides from the DNA; a transport system to separate the single nucleotide from the DNA and incorporate the single nucleotide in a fluorescence-enhancing matrix; and a detection station for the irradiation, detection and identification of the single nucleotides. The nucleotides are advantageously detected by irradiating the nucleotides with a laser to stimulate their natural fluorescence, detecting the fluorescence spectrum and matching the detected spectrum with that previously recorded for the four nucleotides in order to identify the specific nucleotide.
In one embodiment of the Ulmer apparatus an electric field applied (about 0.1–10 V/cm) via suitably incorporated electrodes to induce the chromosomes to migrate into a microchannel single-file, much as is done in an initial step of cell sorting. The individual chromosomes are visualized by the microscope system as they proceed along the microchannel. This step can also be automated by using computer image analysis for the identification of chromosomes (see Zeidler, 1988, Nature 334: 635). Bifurcations in the channel are similarly used in conjunction with selectively applied electric fields to divert the individual chromosomes into small isolation chambers. Once individual chromosomes have been isolated, the sister chromatids are separated by either a focused laser microbeam and optical tweezers, or mechanical microdissection to provide two “identical” copies for sequencing.
The present invention will be seen to use optical tweezers not only on chromosomes and the like once delivered to “chambers” by use of microchannels, but also to divert the particles within the microchannels themselves—a process that Ulmer contemplates to do only by electric fields.
U.S. Pat. No. 5,495,105 to Nishimura, et al. for a METHOD AND APPARATUS FOR PARTICLE MANIPULATION, AND MEASURING APPARATUS UTILIZING THE SAME concerns a flow of liquid containing floating fine particles formed in a flow path, thereby causing successive movement of the particles. A light beam having intensity distribution from a laser is focused on the liquid flow, whereby the particle is optically trapped at the irradiating position, thus being stopped against the liquid flow or being slowed by a braking force. This phenomenon is utilized in controlling the spacing of the particles in the flow or in separating the particles.
The present invention will be seen not to be concerned with retarding (breaking) or trapping the flow of particles in a fluid, but rather in changing the path(s) of particle flow.
The next three patents discussed are not necessarily prior art to the present invention because they have issuance dates that are later than one year prior to the priority date of the present patent application as this priority date is established by the predecessor provisional patent application, referenced above. However, these patents are mentioned for completeness in describing the general current, circa 21001, state of the art in microfluidic and/or laser manipulative particle processing, and so that the distinction of the present invention over existing alternative techniques may be better understood.
In this regard, U.S. Pat. No. 6,139,831 to Shivashankar, et al., assigned to The Rockfeller University (New York, N.Y.), for an APPARATUS AND METHOD FOR IMMOBILIZING MOLECULES ONTO A SUBSTRATE, contemplates both (i) movement by microfluidics and (ii) manipulation by optical tweezers. However, Shivashankar, et al. contemplate that these should be separate events.
The Shivashankar, et al., patent concerns an apparatus and method for immobilizing molecules, particularly biomolecules such as DNA, RNA, proteins, lipids, carbohydrates, or hormones onto a substrate such as glass or silica. Patterns of immobilization can be made resulting in addressable, discrete arrays of molecules on a substrate, having applications in bioelectronics, DNA hybridization assays, drug assays, etc. The Shivashankar, et al., invention reportedly readily permits grafting arrays of genomic DNA and proteins for real-time process monitoring based on DNA-DNA, DNA-protein or receptor-ligand interactions. In the apparatus an optical tweezer is usable as a non-invasive tool, permitting a particle coated with a molecule, such as a bio-molecule, to be selected and grafted onto spatially localized positions of a semiconductor substrate. It is recognized that this non-invasive optical method, in addition to biochip fabrication, has applications in grafting arrays of specific biomolecules within microfluidic chambers, and it is forecast by Shivashankar, et al., that optical separation methods may work for molecules as well as cells.
Well they may; however the present invention will be seen, inter alia, to employ optical tweezers on biomolecules while moving these molecules move in microchannels under microfluidic forces—as opposed to being stationary in microfluidic chambers.
U.S. Pat. No. 6,159,749 to Liu, assigned to Beckman Coulter, Inc. (Fullerton, Calif.), for a HIGHLY SENSITIVE BEAD-BASED MULTI-ANALYTE ASSAY SYSTEM USING OPTICAL TWEEZERS concerns an apparatus and method for chemical and biological analysis, the apparatus having an optical trapping means to manipulate the reaction substrate, and a measurement means. The optical trapping means is essentially a laser source capable of emitting a beam of suitable wavelength (e.g., Nd:YAG laser). The laser beam impinges upon a dielectric microparticle (e.g., a 5 micron polystyrene bead which serves as a reaction substrate), and the bead is thus confined at the focus of the laser beam by a radial component of the gradient force. Once “trapped,” the bead can be moved, either by moving the beam focus, or by moving the reaction chamber. In this manner, the bead can be transferred among separate reaction wells connected by microchannels to permit reactions with the reagent affixed to the bead, and the reagents contained in the individual wells.
The patent of Liu thus describes the act of moving particles—beads—in microchannels under force of optical laser beams, or traps. However, as with the other references, Liu does not contemplate that particles moving under force of microfluidics should also be subject to optical forces.
U.S. Pat. No. 6,294,063 to Becker, et al., assigned to the Board of Regents, The University of Texas System (Austin, Tex.), for a METHOD AND APPARATUS FOR PROGRAMMABLE FLUIDIC PROCESSING concerns a method and apparatus for microfluidic processing by programmably manipulating a packet. A material is introduced onto a reaction surface and compartmentalized to form a packet. A position of the packet is sensed with a position sensor. A programmable manipulation force is applied to the packet at the position. The programmable manipulation force is adjustable according to packet position by a controller. The packet is programmably moved according to the programmable manipulation force along arbitrarily chosen paths.
It is contemplated that the “packets” may be moved along the “paths” by many different types of forces including optical forces. The forces are described to be any of dielectrophoretic, electrophoretic, optical (as may arise, for example, through the use of optical tweezers), mechanical (as may arise, for example, from elastic traveling waves or from acoustic waves), or any other suitable type of force (or combination thereof). Then, in at least some embodiments, these forces are programmable. Using such programmable forces, packets may be manipulated along arbitrarily chosen paths.
As with the other described patents, the method and apparatus of Becker, et al., does not contemplate moving with one force—microfluidics—while manipulating with another force—an optical force.